Methods of treating muscle cramping and related compositions

ABSTRACT

Described herein are compositions comprising capsaicin and methods of treating muscle cramping in a subject, comprising orally administering to the subject a composition comprising capsaicin and an excipient.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a division of U.S. application Ser. No. 14/450,384,filed Aug. 4, 2014, which is a continuation of U.S. application Ser. No.13/191,941, filed Jul. 27, 2011, now abandoned, which claims the benefitof U.S. Provisional Application No. 61/368,059, filed Jul. 27, 2010. Thecontents of each of the foregoing applications are hereby incorporatedby reference in their entirety.

BACKGROUND OF THE INVENTION

In general, the invention relates to methods and compositions forpreventing, treating or ameliorating muscle cramping and/or acceleratingnerve-muscle recovery from exercise fatigue.

Muscle cramps, the involuntary and forceful contraction of muscles, areoften painful and can last for a prolonged period of time. Musclecontractions and cramping can be triggered by exercise and can alsooccur spontaneously (e.g., nocturnal or night cramps). The underlyingphysiological mechanism of muscle cramping is unknown. Recentunderstanding has led to the hypothesis that cramping results fromexcessive electrical firing of the neurons (motor neurons) that projectfrom the spinal cord and trigger contraction of skeletal muscles(Schwellnus, Br J Sports Med. 43:401-8, 2009; Miller et al, Med SciSports Exerc. 42:953-61, 2010). Recovery from strenuous exercise can beassociated with neuromuscular irritability, associated withneuromuscular fatigue, that may or may not be associated with thedevelopment of frank cramps. Few treatments and therapeutic regimens areavailable to alleviate this neuromuscular irritability.

There exists a need in the art for improved methods and compositions forpreventing, treating, and ameliorating muscle cramping and/oraccelerating nerve-muscle recovery from exercise fatigue by reducingneuromuscular irritability. As shown herein, compositions that includeactivators of TRP and ASIC channels may be useful to prevent, treat, orameliorate muscle cramping and/or accelerate nerve-muscle from, e.g.,exercise fatigue. Further, these compositions can be useful in treatingneuromuscular irritability that may or may not be associated with thedevelopment of frank cramps.

SUMMARY OF THE INVENTION

The present invention is directed to the prevention, treatment oramelioration of muscle cramps and/or accelerating nerve-muscle recoveryfrom exercise fatigue using a composition with an activator of TRPV1channels, an activator of TRPA1 channels, and/or an activator of ASICchannels.

In a first aspect, the invention features a method for treating musclecramps in a subject in need thereof (e.g., a human), the methodincluding the step of administering to the subject a composition thatincludes an effective amount of one or more TRPV1 channel activators,TRPA1 channel activators, or ASIC channel activators, or any combinationthereof.

The invention also features a method for treating musculoskeletalirritability in a subject in need thereof. The method includesadministering to the subject a composition that includes an effectiveamount of one or more TRPV1 channel activators, TRPA1 channelactivators, or ASIC channel activators, or any combination thereof.

In still another aspect, the invention features a method for improvingmuscle recovery (e.g., muscle recovery following exercise) in a subjectin need thereof, where the method includes administering to the subjecta composition that includes an effective amount of one or more TRPV1channel activators, TRPA1 channel activators, or ASIC channelactivators, or any combination thereof.

In any of the foregoing methods, the composition can be, e.g., an oralformulation (e.g., a liquid, beverage, gel, semi-solid, frozen liquid,lozenge, hard candy, dissolving strip, or spray).

In certain embodiments of any of the methods described herein, thecomposition is administered to the subject prior to exercise, duringexercise, or following exercise (e.g., within 0-120 minutes prior toexercise, or within 0-360 minutes, 0-15 minutes, or 2-6 hours followingexercise.

Muscle cramps that can be treated or prevented using the methods andcompositions described herein include, e.g., muscle cramps resultingfrom exercise, nocturnal cramps, and menstrual cramps.

In particular embodiments of any of the methods described herein, thecomposition includes an effective amount of two or three different TRPchannel activators independently selected from:

(a) capsaicin or other capsaicinoids;

(b) cinnamaldehyde or cinnamon oil; and

(c) gingerols.

In other embodiments of any of the methods described herein, eachchannel activator, independently, includes between 0.001% to 1% weightpercent of a composition that is a solid, semi-solid, gel, or chewinggum, or 0.001 to 1% (v/v) of a composition that is a liquid, beverage,or spray.

In still other embodiments of any of the methods described herein, thecomposition is any of the compositions described herein.

The invention also features a composition formulated for oral ingestionby a subject. The composition includes an effective amount of one ormore channel activators (e.g., one or more substantially pure channelactivators) selected from TRPV1 channel activators, TRPA1 channelactivators, and ASIC channel activators. Desirably, the composition is aliquid, beverage, gel, semi-solid, frozen liquid, lozenge, hard candy,dissolving strip, or spray.

Desirably, the channel activator is capable of activation of a channelin a gastroesophogeal neuron when administered to a subject

The channel activators can be substantially pure or not substantiallypure (e.g., part of a crude extract).

In certain embodiments, the channel activators are capable of activationof a channel in a gastroesophogeal neuron when administered to asubject.

The TRPV1 channel activator can be is a capsaicinoid (e.g., capsaicin).In one example embodiments, the composition contains capsaicin but issubstantially free of dihydrocapsaicin.

The TRPV1 channel activator can also be, e.g., oleoylethanolamide,N-oleoyldopamine, 3-methyl-N-oleoyldopamine, oleamide, capsiate, a1-monoacylglycerol having C18 and C20 unsaturated and C8-C12 saturatedfatty acid, a 2-monoacylglycerol having C18 and C20 unsaturated fattyacids, miogadial, miogatrial, polygodial, a terpenoid with analpha,beta-unsaturated 1,4-dialdehyde moiety, sanshool, evodiamine,acesulfame-K, cyclamate, CuSO₄, ZnSO₄, FeSO₄, arvanil, anandamide,N-arachidonoyl-dopamine, flufenamic acid dopamide, a dopamine amide offenamic acid, 4-hydroxynonenal, or1-[2-(1-adamantyl)ethyl]-1-pentyl-3-[3-(4-pyridyl)propyl]urea, orgingerol.

Suitable TRPA1 channel activators include, e.g., allyl isothiocyanate,gingerols, cinnamaldehyde, acrolein, farnesyl thiosalicylic acid,A-tetrahydrocannabinol, eugenol, shogaols, nicotine, nicotinederivatives or analogs, methyl salicylate, cinnamaldehyde, allicin,diallyl sulfide, diallyl disulfide, diallyl trisulfide, sanshools, andfarnesyl thioacetic acid.

Each channel activator can be present, e.g., between 0.001% to 1% (w/w)or 0.001 to 1% (v/v) of a composition.

In certain embodiments, the composition includes no more than one TRPV1channel activator, TRPA1 channel activator, or ASIC channel activator.In still other embodiments, the composition includes a TRPV1 channelactivator and a TRPA1 channel activator (e.g., a substantially pureTRPA1 channel activator and/or a substantially pure TRPV1 channelactivator). In yet other embodiments, the composition includes no morethan one TRPV1 channel activator and no more than one TRPA1 channelactivator. And in still other embodiments, the composition includes asecond TRPV1 channel activator and/or a second TRPA1 channel activator.

The composition includes can include an effective amount of an ASICchannel activator (e.g., a substantially pure ASIC channel activator).In some embodiments, the ASIC channel activator (e.g., a substantiallypure ASIC channel activator) is capable of activation of an ASIC channelin a gastroesophogeal neuron when administered to a subject.

In certain embodiments, the ASIC channel activator is an acidulant thatis acetic acid. In further embodiments, the composition is a liquid orbeverage that has an acetic acid concentration ranging from 0 M to 0.1 M(e.g., from 0 M to 0.001 M). In other embodiments, the ASIC channelactivator is an acidulant selected from phosphoric acid, citric acid,malic acid, succinic acid, tartaric acid, fumaric acid, and ascorbicacid.

The invention also features a composition that includes an effectiveamount of two or three different TRP channel activators independentlyselected from capsaicin or another capsaicinoid; cinnamaldehyde orcinnamon oil; and gingerols, where the composition is an oralformulation that is a liquid, beverage, gel, solid, semi-solid, chewinggum, or spray.

In certain embodiments, the composition includes an effective amount oftwo or three different TRP channel activators independently selectedfrom: capsicum; cinnamon volatile oil; and ginger oleoresin. Forexample, the the composition can include each of the components.

In some embodiments the capsaicinoid is present in 0.001% to 1% (w/w) or0.001% to 1% (v/v); and/or the cinnamaldehyde or cinnamon oil is presentin 0.001% to 10% (w/w) or 0.001% to 10% (v/v); and/or the gingerols arepresent in 0.001% to 10% (w/w) or 0.001% to 10% (v/v).

In one example, the composition is a beverage that optionally includes asweetener.

A composition of the invention can also include an acidulant selectedfrom acetic acid, phosphoric acid, citric acid, malic acid, succinicacid, tartaric acid, fumaric acid, acetic acid, and ascorbic acid.

The composition can also include a potassium salt at a concentration ofbetween about 0.02% and about 7% by weight based on total volume of theliquid, beverage, or gel.

The composition can be a liquid, beverage, or gel that also includes aviscosity modifier, such as collagen, gellan gum, carbohydrategel-forming polymers, carob bean gum, locust bean gum, carrageenan,alginic acid, sodium alginate, potassium alginate, ammonium alginate,calcium alginate, agar, guar gum, xanthan gum, carboxymethyl cellulose,clear starch, pectin, gelatin, arrowroot, cornstarch, katakuri starch,potato starch, sago, tapioca, furcellaran, or sodium pyrophosphate. Incertain embodiments, the composition has a viscosity between about 1000and about 10000 cP (e.g., 1000, 2000, 3000, 4000, 5000, 6000, 7000,8000, 9000, or 10000 cP).

In some embodiments of any compositions described herein, thecomposition also includes one or more of electrolytes, sweeteners,flavoring agents, vitamins, minerals, amino acids, or preservatives.

In certain embodiments of any compositions described herein, thecomposition is a beverage or gel that is made by reconstituting a drypowder with an aqueous fluid (e.g., water).

In other embodiments of any compositions described herein, thecomposition is a packaged beverage. In some embodiments, the packagedbeverage is provided in a unit that contains between 10-1000 mL (e.g.,between 10-500 mL) of the beverage.

In still other embodiments of any compositions described herein, thecomposition is a gel. In certain embodiments, the composition is apackaged gel. In further embodiments, the packaged gel is provided in aunit that contains between 5-100 grams (e.g., between 30-40 grams) ofthe gel.

In some embodiments of any of the methods described herein, thecomposition of (1) or (2) has a pH that is greater than about 2.5 (e.g.,the pH of the composition is greater than about 2.5, 2.6, 2.7, 2.8, 2.9,3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3,4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, or 6.5). For example, the compositionof (1) or (2) can have a pH of about 4 to about 9, about 5 to about 9,or 6.5 to about 7.5.

In certain embodiments, the composition is a liquid or beverage thatdoes not include acetic acid or a salt thereof, or that has an aceticacid concentration ranging from 0 M to 0.1 M, 0 M to 0.001 M, issubstantially 0 M.

In some embodiments of any of the compositions described herein, theTRPV1 channel activator is a proton concentration of 10⁻⁷ M to 10⁻² M(pH 2-pH 7), e.g. produced by inclusion of acidulants such as aceticacid, phosphoric acid, citric acid, malic acid, succinic acid, tartaricacid, fumaric acid, and ascorbic acid.

In some embodiments of any of the methods or compositions describedherein, where the composition includes vinegar (e.g., apple cidervinegar), the composition includes no more than one of garlic (e.g.,garlic extracts) or ginger (e.g., ginger extracts).

In other embodiments of any of the methods or compositions describedherein, where the composition includes garlic (e.g., garlic extracts),the composition includes no more than one of vinegar (e.g., apple cidervinegar) or ginger (e.g., ginger extracts).

In some embodiments of any of the methods or compositions describedherein, where the composition includes ginger (e.g., ginger extracts),the composition includes no more than one of garlic (e.g., garlicextracts) or vinegar (e.g., apple cider vinegar).

Any of the compositions described herein can be used in any of themethods described herein (e.g., to treat muscle cramps such as nocturnalcramps or menstrual cramps, or muscle cramps resulting from exercise, totreat musculoskeletal irritability, or to improve muscle recovery). Thecompositions used in the methods can include substantially pure channelactivators. In other embodiments of any of the compositions or methodsdescribed herein, the TRP channel (e.g., a TRPV1 or TRPA1 activator)activator and/or the ASIC channel activator is not substantially pure.For example, a composition can include a mixture of capsaicinoids (e.g.,as capsicum or between 0.0001-0.01 mgs/mL of total capsaicinoids). Insome embodiments, the channel activators are provided as extractssuitable for human consumption.

In still another aspect; the invention features a method of preparingany of the compositions described herein for treating or amelioratingmuscle cramps in a subject in need thereof, or for treatingmusculoskeletal irritability in a subject in need thereof, the methodincluding the use of TRP or ASIC channel activator compounds that aresubstantially pure with an excipient to provide a composition that is aliquid, beverage, gel, solid, semi-solid, chewing gum, or spray.

By “ASIC channel” is meant an acid sensing ion channel that is opened bylow pH and can excite certain neurons or muscle fibers or other cells.

By “acidulant” is meant an acidic compound (e.g., citric acid) used tolower the pH of a composition, e.g., the pH can be lowered in the rangeof 2.5-6.5 (e.g., pH of 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, or 6.5).

By “activator,” “compound that activates,” or “agonist” is meant amolecule that stimulates a biological response.

By “administering” and “administration” is meant a mode of delivery. Adaily dosage can be divided into one, two, three or more doses in asuitable form to be administered one, two, three or more timesthroughout a time period. In preferred embodiments of the presentinvention, compositions and solutions are administered orally.

By “beverage” is meant a composition that is not in solid or gas form,such as a liquid or semi-liquid that is designed to enter into the mouthof a subject and be orally consumed or ingested. A beverage may be in aready-to-drink liquid form (e.g., may be consumed without modification)or in a liquid, solid, or concentrated form, which can be transformedinto a ready-to-drink liquid form with an addition of another liquid(e.g., water). See, e.g., Sports Drinks: Basic Science and PracticalAspects, Ed. Ronald J. Maughan, CRC Press, 2000.

Where the term “composition” is used to describe a formulation thatincludes an activator of TRPV1 channels, and/or an activator of TRPA1channels, and/or an activator of ASIC channels, the term refers to acomestible formulation that is suitable for oral ingestion by thesubject (e.g., the human subject). Exemplary compositions that includean activator of TRPV1, TRPA1, and ASIC channels include sprays (e.g.,aerosols), powders, chewing gum, ingestible solids, gels, aqueousbeverages, dry powder (e.g., a powder that can be directly consumed orthat can be reconstituted with liquid to provide a beverage as definedherein), nutritional bars, lozenges, tablets, capsules, wafers, pastes,and the like. Other compositions are described herein.

The term an “effective amount” of a compound as used herein, is thatamount sufficient to effect beneficial or desired results, such as theeffective treatment of muscle cramps or musculoskeletal irritability orthe improvement of muscle recovery following exercise, and, as such, an“effective amount” depends upon the context in which it is beingapplied. For example, in the context of administering an agent thatactivates a TRP channel (e.g., TRPV1 or TRPA1) or an ASIC channel, aneffective amount of an agent is, for example, an amount sufficient toachieve an increase in TRPV1, TRPA1, or ASIC activity as compared to theresponse obtained without administration of the agent. The effectiveamount of active compound(s) used to practice the present invention canalso be varied based on, for example, the age, and body weight, of thesubject or the nature of the exercise.

The compositions can also include one more excipients that are notactivators of TRPV1, TRPA1, and ASIC channels and that are non-toxic andnon-inflammatory in a subject (e.g., in a human subject). In someembodiments, the excipient(s) can provide desirable or improved physicaland/or chemical properties such as stability, flow, viscosity, rate ofdisintegration, taste, delivery, etc. Exemplary, non-limiting excipientsthat can be selected from: a disintegrant (e.g., carmellose, starch,crystalline cellulose, low-substituted hydroxypropyl cellulose, and thelike), a binder (e.g., gum acacia, carmellose, gelatin, crystallinecellulose, simple syrup, honey, hydroxypropyl cellulose, povidone,methylcellulose, and the like), a surfactant (e.g., polyoxyl 40stearate, polysorbate 80, polyoxyethylene hydrogenated castor oil, andthe like), an emulsifier (e.g., polyoxyl 40 stearate, sorbitansesquioleate, polysorbate 80, sodium lauryl sulfate, lauromacrogol, gumarabic, cholesterol, stearic acid, povidone, glyceryl monostearate, andthe like), a plasticizer (e.g., glycerin, propylene glycol, macrogol,and the like), a lubricant (e.g., magnesium silicate, carmellose, lightanhydrous silicic acid, stearic acid, calcium stearate, magnesiumstearate, talc, and the like), a sweetener (e.g., white soft sugar,honey, simple syrup, glucose, saccharin sodium, acesulfame potassium,disodium glycyrrhizinate, and the like), a pH-adjusting agent (e.g.,hydrochloric acid, citric acid, sodium hydrogen carbonate, potassiumhydroxide, sodium hydroxide, sodium carbonate, and the like), apreservative (e.g., benzoic acid, benzalkonium chloride, ethylparahydroxybenzoate, butyl parahydroxybenzoate, propylparahydroxybenzoate, methyl parahydroxybenzoate, and the like), a flavor(e.g., fennel oil, orange oil, cinnamon oil, thymol, orange peeltincture, dl-menthol, 1-menthol, eucalyptus oil, and the like), or acoloring agent (e.g., Food Red No. 2, No. 3, No. 40, No. 102, No. 104,No. 105 or No. 106, Food Yellow No. 4 or No. 5, Food Green No. 3, FoodBlue No. 1 or No. 2, titanium dioxide, sodium copper chlorophyllin,turmeric, gardenia, annatto dye, kaoliang dye, and the like), or anantioxidant (e.g., ascorbic acid, sodium thiosulfate, tocopherol, sodiumhydrogen sulfite, and the like), or any combination thereof.

By “ingestible solid” is meant a solid formulation that can be ingestedby a subject (e.g., a human) without toxic effects.

By “muscle cramp” is meant a spontaneous contraction of one or moremuscles. A muscle cramp may be associated with strenuous exercise orfatigue or may occur during rest (e.g., a nocturnal cramp). Menstrualcramps are also muscle cramps By “nerve-muscle recovery” or “musclerecovery” is meant the recovery from spontaneous muscle contractions andfatigue following exercise that may or may not be associated with thedevelopment of frank cramps.

By “neuromuscular irritability” is meant spontaneous muscle contractions(e.g. associated with muscle fatigue) which may or may not be associatedwith frank cramps.

By “preventing” or “reducing the likelihood of” is meant reducing theseverity, the frequency, and/or the duration of a condition or disorder(e.g., muscle cramping) or the symptoms thereof. For example, reducingthe likelihood of or preventing muscle cramping is synonymous withprophylaxis of muscle cramping.

By “subject” is meant a mammal, including, but not limited to, a humanor non-human mammal, such as a bovine, equine, canine, ovine, or feline.

As used herein, and when used in reference to TRPA1, TRPV1, and/or ASICchannel activators, the term “substantially pure” refers to acomposition that includes a channel activator in which the compositionis free of organic and/or inorganic species that do not activate theTRPA1, TRPV1, and/or ASIC channels, and where 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, 97%, 98%, 99%, or 99.5% (w/w) of the composition is aparticular channel activator compound. Substantially pure compositionscan be prepared and analyzed using standard methods known in the art(e.g., chromatographic separation, extractions, and the like).Substantially pure compositions can include isomeric impurities (e.g.,geometric isomers) and/or salts or solvates of a channel activator.

By “treating” or “ameliorating” is meant administering a composition fortherapeutic purposes or administering treatment to a subject alreadysuffering from a disorder to improve the subject's condition. By“treating a condition or disorder” or “ameliorating a condition ordisorder” is meant that the condition or disorder (e.g., musclecramping) and the symptoms associated with the condition or disorderare, e.g., alleviated, reduced, cured, or placed in a state ofremission. As compared with an equivalent untreated control, suchamelioration or degree of treatment is at least 5%, 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100%, as measured by any standardtechnique.

By “transient receptor potential cation channel, subfamily V, member 1”or “TRPV1” is meant a nonselective cation channel that may be activatedby physical and/or chemical stimuli. TRPV1 channel activators include,e.g., capsaicinoids and capsaicinoid analogs or derivatives and certainendocannabinoids.

By “transient receptor potential cation channel, subfamily A, member 1”or “TRPA1” is meant a cation channel that is a member of the transientreceptor potential channel family. TRPA1 channel activators include,e.g., ally isothiocyanate, cinnamaldehyde, farnesyl thiosalicylic acid,nicotine and its structural analogues, formalin, hydrogen peroxide,4-hydroxynonenal, and acrolein.

By “viscosity” is meant a measurement of a fluid's internal resistanceto flow (e.g., “thickness”). Viscosity is generally expressed incentipoise (cP) or pascal-seconds.

Other features and advantages of the invention will be apparent from thedetailed description and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows graphs from 6 sensory neurons isolated from the trigeminalganglia of rats that illustrate their activation by the capsicum,cinnamon, and ginger extracts that were used in the human experiments.These data show that each agent is capable of acting alone to activatesome neurons but also that a combination of agents can produce strongeractivation of a larger fraction of neurons. Further, the bottom tworecords suggest that there can be strongly synergistic activation ofneurons by the capsicum extract and the ginger extract when applied incombination.

FIG. 2 shows the effect of the TRP-Stim beverage on cramping of theflexor hallucis brevis (FHB) of Subject A. Under control conditions,cramping was reliably induced by stimulating the muscle using anelectrical muscle stimulator. After ingestion of 50 mL of a TRP-Stimbeverage containing capsaicin and capsaicinoids (TRPV1 agonists),cinnamaldehyde (TRPA1 agonist), and gingerols (TRPA1 and TRPV1agonists), cramping was very brief after 11 minutes and essentiallyabsent at tests at 20 minutes and 2½ hours after ingestion.

FIG. 3 shows the effect of the TRP-Stim beverage on cramping of the FHBof a second subject after cramping was induced. In recordings beginning12 minutes after ingestion of the TRP-Stim beverage, stimulation at 10Hz or 12 Hz produced essentially no cramping, and increasing thefrequency of stimulation to 14 Hz also did not induce cramping. Thedramatic reduction in cramping was still present 4 hours later in thissubject.

FIG. 4 shows the effect of the TRP-Stim beverage on cramping of the FHBof a third subject tested over longer times. Under control conditions, acramp lasting 58 seconds was induced. After ingestion of the TRP-Stimbeverage, the duration of the cramp was reduced to 27 seconds after 8minutes, to 8 seconds after 15 minutes, and cramping was abolished after20 minutes and in a test after 2 hours. In tests 11 hours afteringestion, reliable cramping had returned. After the subject again drank50 mL of the TRP-Stim beverage, cramping was completely abolished intests beginning after 10 minutes.

FIG. 5 shows the effect of the TRP-Stim beverage on cramping of the FHBof a fourth subject. This subject had engaged in strenuous exercise(triathlon) four hours earlier and was experiencing muscle twitchiness.This subject had an unusually low frequency threshold (8 Hz) forinduction of cramping in the FHB muscle, and the resulting cramps wereunusually long (172 seconds after 8 Hz stimulation and 222 seconds after10 Hz stimulation). Cramping was completely gone in tests starting 13minutes after ingestion of the TRP-Stim beverage, even when increasingthe stimulation frequency to 12 Hz. Cramping was still abolished 3 hourslater. After 4 hours, cramping returned, but with an increased frequencythreshold (10 Hz). After the subject again drank 50 mL of the TRP-Stimbeverage, cramping was again completely abolished.

FIG. 6 is a graph showing the effect of the TRP-Stim beverage oncramping of the gastroenemius (calf) muscle of a fifth subject. Themuscle was stimulated, and after cessation of stimulation, the musclewent into a prolonged cramp lasting 59 seconds. In a test 3 minutesafter ingestion of 50 mL of TRP-Stim, cramping was abolished.

FIG. 7 shows the effect of the TRP-Stim beverage on cramping of thegastroenemius (calf) muscle of a sixth subject. The muscle wasstimulated, and after cessation of stimulation, the muscle went into aprolonged cramp lasting 96 seconds. In a test 4 minutes after ingestionof 50 mL of TRP-Stim, cramping was abolished. Cramping was stillabolished in a test conducted 40 minutes later.

FIG. 8 is a graph showing the effect of the TRP-Stim beverage oncramping of an FHB muscle in a seventh subject who experiencedspontaneous cramping induced by pointing her toe. Ten minutes after thesubject ingested 50 mL of the TRP-Stim beverage, cramping was abolished.

DETAILED DESCRIPTION

The methods and compositions of the present invention are directed tothe treatment or amelioration of muscle cramps using a composition thatincludes one or more TRPV1 channel activators and/or TRPA1 channelactivators and/or ASIC channel activators.

Muscle Cramping

Few treatments and therapeutic regimens are available to alleviatemuscle cramping and associated symptoms. Recent work suggests thatingestion of pickle juice can alleviate cramping (Miller et al., Med.Sci. Sports. Exerc. 42:953-61, 2010). The mechanism for the efficacy ofpickle juice is unknown, although it has been recently proposed (Milleret al., Med Sci Sports Exerc. 42:953-61, 2010) that it involves anoropharangeal reflex induced by the sour taste of acetic acidstimulating taste receptors (Kajii et al. Physiol Behav. 77:321-5,2002). Without being bound by theory, we instead hypothesize that picklejuice's efficacy results from the activation of a specific class ofnon-taste primary sensory neurons that contain nerve endings in themouth, esophagus, and stomach. These neurons, polymodal C-fiber andA-delta neurons, are distinct from taste neurons, are activated bydiverse (polymodal) stimuli, including noxious mechanical, chemical, andthermal stimuli, and are known to contain specific ion channel receptorscalled Transient Receptor Potential (TRP) ion channels and acid sensing(ASIC) ion channels (Beilefeldt et al., Am J Physiol Gastrointest LiverPhysiol 294: G130-G138, 2008; Yu et al., Am J Physiol Gastrointest LiverPhysiol 297: G34-G42, 2009).

We hypothesize that cramps result from excessive firing of motor neuronsin the spinal cord, in accordance with recent understanding (Schwellnus,Br J Sports Med. 43:401-8, 2009). We further hypothesize thatstimulating TRPV1 or TRPA1 or ASIC channels in the nerve endings ofprimary sensory neurons present in the mouth, esophagus and/or stomach,which project to the spinal cord and brain stem, influence neuromuscularactivity by altering activity of neural circuits in the spinal cord orbrainstem. TRPV1, TRPA1, and ASIC channels are known to be present ingastroesophageal polymodal sensory neurons that mediate transmission ofa variety of noxious stimuli, including mechanical, chemical, andthermal stimul (Beilefeldt et al., Am J Physiol Gastrointest LiverPhysiol 294: G130-G138, 2008; Yu et al., Am J Physiol Gastrointest LiverPhysiol 297: G34-G42, 2009). These primary sensory neurons project tothe spinal cord and brain stem, where they release glutamate and avariety of neuropeptides (e.g., calcitonin gene-related peptide (CGRP)and substance P). These transmitters act broadly on other types ofneurons within the spinal cord and brain stem circuitry, includingneurons that release GABA, glycine, and serotonin in the spinal cord,which can in turn inhibit firing of motor neurons. Activity ofinhibitory neurons in the spinal cord and brain stem may thus betriggered by stimulation of molecular targets (e.g., TRPV1 and TRPA1 andASIC channels) present in primary sensory nerve endings in the mouth,esophagus and/or stomach, resulting in inhibition of the motor neuronswhose excessive firing is responsible for muscle cramping. In support ofthis hypothesis, some cramps can be prevented by local nerve block andappear to result from impairment of function of GABAergic interneuronsin the spinal cord (Obi et al., Muscle and Nerve 6:1228-1231, 1993).

We also hypothesize that spontaneous muscle contractions during recoveryfrom exercise fatigue also arise from excessive firing of motor neuronsin the spinal cord and can similarly be reduced by stimulating TRPV1,TRPA1, and/or ASIC channels in the nerve endings of primary sensoryneurons present in the mouth, esophagus and/or stomach, which project tothe spinal cord and brain stem. By reducing spontaneous musclecontractions stimulation of TRPV1, TRPA1, and/or ASIC channels in nerveendings within the esophagus and/or stomach can accelerate neuromuscularrecovery following exercise fatigue.

Compositions

The compositions described herein are comestible formulations suitablefor oral consumption by a subject (e.g., by a human) and include one ormore activators of TRPV1, TRPA1, and/or ASIC channels as well as one ormore optional excipients as described herein. Exemplary, non-limitingcompositions include those that are solids (e.g., chews or chewinggums), liquids (e.g., beverages), and gels.

TRPV1 Channel Activators

Compounds that activate TRPV1 that may be used in the compositions ofthe present invention include, for example, capsaicin, capsaicin analogsand derivatives (e.g., capsaicinoids), and any other compound thatactivates TRPV1, examples of which are described herein. Modulators ofTRPV1 activity are known in the art (see, e.g., Harteneck et al.,“Synthetic modulators of TRP channel activity,” Adv Exp Med Biol.704:87-106, 2011, and other references described herein).

In one embodiment, the TRPV1 channel activator is a capsaicinoid (e.g.,capsaicin (8-methyl-N-vanillyl-trans-6-nonenamide)). Exemplarycapsaicinoids are provided in Table 1.

TABLE 1 Exemplary capsaicinoids Capsaicin

Dihydrocapsaicin

Nordihydrocapsaicin

Homodihydrocapsaicin

Homocapsaicin

Nonivamide

Suitable capsaicinoids and capsaicinoid analogs and derivatives for usein the compositions and methods of the present invention includenaturally occurring and synthetic capsaicin derivatives and analogsincluding, e.g., vanilloids (e.g., N-vanillyl-alkanedienamides,N-vanillyl-alkanedienyls, and N-vanillyl-cis-monounsaturatedalkenamides), capsiate, dihydrocapsiate, nordihydrocapsiate and othercapsinoids, capsiconiate, dihydrocapsiconiate and other coniferylesters, capsiconinoid, resiniferatoxin, tinyatoxin, civamide,N-phenylmethylalkenamide capsaicin derivatives, olvanil,N-[(4-(2-aminoethoxy)-3-methoxyphenyl)methyl]-9Z-octa-decanamide,N-oleyl-homovanillamide, triprenylphenols (e.g., scutigeral), gingerols,piperines, shogaols, guaiacol, eugenol, zingerone, nuvanil, NE-19550,NE-21610, and NE-28345.

Other suitable TRPV1 channel activators include oleoylethanolamide,N-oleoyldopamine, 3-methyl-N-oleoyldopamine, oleamide, capsiate,1-monoacylglycerols having C18 and C20 unsaturated and C8-C12 saturatedfatty acid, 2-monoacylglycerols having C18 and C20 unsaturated fattyacids, miogadial, miogatrial, polygodial, and other terpenoids with analpha,beta-unsaturated 1,4-dialdehyde moiety, sanshools, evodiamine,acesulfame-K, cyclamate, sulfates (e.g., CuSO₄, ZnSO₄, and FeSO₄),arvanil, anandamide, N-arachidonoyl-dopamine, flufenamic acid dopamideand other dopamine amides of fenamic acids, 4-hydroxynonenal, SA 13353(i.e., 1-[2-(1-adamantyl)ethyl]-1-pentyl-3-[3-(4-pyridyl)propyl]urea),gingerol or salts of magnesium.

In addition, the TRPV1 channel activator may be an analog or derivativeof any of the TRPV1 channel activators described herein.

Additional TRPV1 channel activators are described, for example, in U.S.Pat. Nos. 7,632,519; 7,446,226; 7,429,673; 7,407,950; 6,022,718;5,962,532; 5,762,963; 5,403,868; 5,290,816; 5,221,692; 4,812,446;4,599,342; 4,564,633; 4,544,669; 4,544,668; 4,532,139; 4,493,848;4,424,205; 4,313,958; in U.S. Patent Application Publication Nos.2007/0293703; 2007/0167524; 2006/0240097; and 2005/0085652; and in WO00/50387, each of which is incorporated by reference.

In addition, the TRPV1 channel activator may be an acidulant (e.g.,acetic acid, phosphoric acid, citric acid, malic acid, succinic acid,tartaric acid, fumaric acid, or ascorbic acid) maintaining a low pH inthe range of 2.5-6.5 (e.g., pH of 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5,6.0, or 6.5).

TRPV1 channel activators for use in the compositions and methodsdescribed herein can be identified using standard methodology, asdescribed, for example, in U.S. Patent Application Publication No.2003/0104085, which is hereby incorporated by reference. Exemplaryassays for identification of TRPV1 channel activators include, withoutlimitation, receptor binding assays; functional assessments ofstimulation of calcium influx or membrane potential in cells expressingthe TRPV1 receptor; assays for the ability to induce cell death in suchcells (e.g., selective ablation of C-fiber neurons); and other assaysknown in the art.

A TRPV1 channel activator may be present in a composition of theinvention at a concentration range of about 0.01% to 10% by weight byweight based on the total volume of the composition (e.g., 0.01, 0.1,0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%), though a TRPV1 channelactivator may be present in lower or higher concentrations.

TRPA1 Channel Activators

TRPA1 channels are activated by naturally occurring substancesincluding, e.g., mustard oil, isothiocyanate compounds (e.g., allylisothiocyanate), acrolein, farnesyl thiosalicylic acid.A-tetrahydrocannabinol (THC), eugenol, ginger, gingerol, gingerols,shogaols, nicotine, nicotine derivatives and analogs, methyl salicylate,cinnamaldehyde, cinnamon oil, wintergreen oil, clove oil, allicin,diallyl sulfide, diallyl disulfide, diallyl trisulfide, sanshools,farnesyl thiosalicylic acid, and farnesyl thioacetic acid. The TRPA1channel activator may also be an analog or derivative of any of theTRPA1 channel activators described herein, and additional TRPA1 channelactivators are identified in WO 2009/071631, hereby incorporated byreference. Still other modulators of TRPA1 are described in, e.g.,Harteneck et al., “Synthetic modulators of TRP channel activity,” AdvExp Med Biol. 704:87-106, 2011; Viana et al. “TRPA1 modulators inpreclinical development,” Expert Opin. Ther. Pat. 19(12):1787-99.2009).

Methods for identifying TRPA1 channel activators are known in the artand are described, for example, in U.S. Pat. No. 7,674,594.

A TRPA1 channel activator may be present in a composition of theinvention at a concentration range of about 0.01% to 10% by weight byweight based on the total volume of the composition (e.g., 0.01, 0.1,0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%), though a TRPA1 channelactivator may be present in lower or higher concentrations.

ASIC Channel Activators

ASIC channels are activated by low pH. The pH of a composition of thepresent invention that includes an ASIC channel activator may be in therange of 2.5-6.5 (e.g., pH of 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, or6.5). The pH may be adjusted within this range by any means acceptablefor compositions that are intended to be ingested by a subject.Exemplary acidulants are acetic acid, phosphoric acid, citric acid,malic acid, succinic acid, tartaric acid, fumaric acid, and ascorbicacid. The acidulant may be present in a composition of the invention ata concentration range of about 0.01% to 10% by weight based on the totalvolume of the composition (e.g., 0.01, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8,9, or 10%), though the acidulant may be present in lower or higherconcentrations.

Additional Components of the Composition

The composition of the present invention may additionally include, forexample, electrolytes (e.g., potassium salt or other salts), sweeteners,flavoring and coloring agents, vitamins, minerals, preservatives, andantioxidants.

Viscosity and Viscosity Modifiers

Viscosity is the ratio of shear stress to shear rate, expressed asdynes-second/cm², or poise. A centipoise (cP) is one one-hundredth of apoise.

The composition of the present invention may have a viscosity greaterthan water (i.e., about 1.0 cP at 20° C.), e.g., about 100, 200, 300,400, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 6000,7000, 8000, 9000 cP or more. If a consistency of corn syrup is desired,viscosities in the range of about 2500 cP are suitable. If a consistencyof a soft gel or honey is desired, viscosities in the range of about10000 cP to about 15000 cP are suitable. For pudding-like products,viscosities in the range of about 30000 cP to about 38000 cP aredesirable. Viscosity of the compositions of the present invention may bemeasured with, e.g., a rheometer or viscometer, though additionalmethods of measuring viscosity are known in the art.

Viscosity modifiers may be added to compositions of the presentinvention. Such viscosity modifiers include, for example, collagen,gellan gum, carbohydrate gel-forming polymers, carob bean gum, locustbean gum, carrageenan, alginates (e.g., alginic acid, sodium alginate,potassium alginate, ammonium alginate, and calcium alginate), agar, guargum, xanthan gum, carboxymethyl cellulose, clear starch, pectin,gelatin, arrowroot, cornstarch, katakuri starch, potato starch, sago,tapioca, furcellaran, and sodium pyrophosphate. A viscosity modifier maybe present in the composition in an amount of from about 0.01% to 10% byweight based on the total volume of the composition (e.g., 0.01, 0.1,0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%), though the viscosity modifiermay be present in lower or higher concentrations.

Electrolytes

Exemplary electrolytes include potassium salts, chloride salts, bromidesalts, sodium salts, magnesium salts, calcium salts, citrate salts,acetate salts, phosphate salts, salicylates, bicarbonate salts, lactatesalts, sulphate salts, tartrate salts, benzoate salts, selenite salts,molybdate salts, iodide salts, oxides, and combinations thereof. Anelectrolyte may be present in a composition of the invention at aconcentration range of about 0.01% to 10% by weight based on the totalvolume of the composition (e.g., 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.5,1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%), though an electrolyte may be presentin lower or higher concentrations.

In certain embodiments, the compositions of the present inventioninclude high concentrations of potassium (e.g., potassium chloride). Theconcentration of potassium in the composition may be, e.g., 0.01, 0.02,0.03, 0.04, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, or 7% or more by weightbased on the total volume of the composition.

In certain embodiments, the compositions of the present inventioninclude high concentrations of magnesium (e.g., magnesium chloride). Theconcentration of magnesium in the composition may be, e.g., 0.01, 0.02,0.03, 0.04, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, or 7% or more by weightbased on the total volume of the composition.

Sweeteners

Sweeteners may be included in the compositions of the invention.Exemplary sweeteners include high fructose corn syrup, mannose, maltose,glucose polymers, sucrose (e.g., cane sugar or beet sugar), glucose,dextrose, lactose, galactose, fructose, polysaccharides (e.g.,malodextrins), rice syrup, honey, and natural fruit juices (e.g., orangejuice, papaya juice, pineapple juice, apple juice, grape juice, apricotjuice, pear juice, tomato juice, agave nectar, or cranberry juice).Additionally, non- or low-caloric sweeteners can be used in thecompositions of the invention. Examples of such non-caloric orlow-caloric sweeteners include, but are not limited to, saccharin,cyclamates, acetosulfam, sorbitol, sucralose, xylitol, erythritol,Stevia extract, L-aspartyl-L-phenyl-alanine ester (e.g., aspartame),L-aspartyl-D-alanine alkyl amides,L-aspartyl-L-1-hydroxymethylalkaneamide, andL-aspartyl-1-hydroxyethylalkaneamide. Sweeteners may be present in acomposition of the invention at a concentration range of about 2% to 20%by weight based on the total volume of the composition (e.g., 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20%), thoughsweeteners may be present in lower or higher concentrations.

Flavoring and Coloring Agents

Exemplary flavoring agents include almond oil, amaretto oil, anethole,anise oil, benzaldehyde, blackberry, black walnut oil, blueberry,caraway, caraway oil, cardamom oil, cardamom seed, cherry juice, cherrysyrup, cinnamon, cinnamon oil, cinnamon water, citric acid, citric acidsyrup, clove oil, cocoa, coriander oil, dextrose, eriodictyon, ethylacetate, ethyl vanillin, fennel oil, ginger, glucose, glycerin,glycyrrhiza, grape, honey, lavender oil, lemon oil, lime, mannitol,methyl salicylate, myristica oil, orange oil, orange peel, orange syrup,peppermint, peppermint oil, peppermint water, phenylethyl alcohol,pineapple, raspberry juice, raspberry syrup, rosemary oil, rose oil,rose water, sarsaparilla syrup, sorbitol, spearmint, spearmint oil,strawberry, sucrose, thyme oil, tolu balsam, vanilla, vanillin, and wildcherry syrup. Additional flavoring agents may be found in Food ChemicalsCodex and Fenaroli's Handbook of Flavor Ingredients.

Small amounts of one or more coloring agents may be utilized in thecompositions of the present invention. Coloring agents include, e.g.,beta-carotene, riboflavin dyes, FD&C dyes (e.g., Yellow No. 5, Blue No.1, Blue No. 2, and Red No. 40), FD&C lakes, chlorophylls andchlorophyllins, caramel coloring, annatto, cochineal, turmeric, saffron,paprika, and fruit, vegetable, and/or plant extracts (e.g., grape, blackcurrant, aronia, carrot, beetroot, red cabbage, elderberry, and hibiscusextracts). The amount of coloring agent used will vary depending on theagents used in the composition and the color intensity desired in thefinished product. The amount of coloring agent to be used can be readilydetermined by one skilled in the art.

Vitamins and Minerals

Non-limiting examples of vitamins and minerals that may be included inthe compositions of the present invention include, e.g., cholinebitartate, niacinamide, thiamin, folic acid, d-calcium pantothenate,biotin, vitamin A, vitamin C, vitamin B₁ hydrochloride, vitamin B₂,vitamin B₃, vitamin B₆ hydrochloride, vitamin B₁₂, vitamin D, vitamin Eacetate, vitamin K, and salts of calcium, potassium, magnesium, zinc,iodine, iron, and copper. When included in a composition of theinvention, the composition contains at least 5, 10, 15, 20, 25, 30, 35,40, 45, or 50% of the U.S. recommended daily intake (RDI) for suchvitamins and minerals.

Preservatives

One or more preservatives may additionally be utilized in thecompositions described herein. Exemplary preservatives include, forexample, sorbate, benzoate, and polyphosphate preservatives (e.g.,sorbic acid, benzoic acid, calcium sorbate, sodium sorbate, potassiumsorbate, calcium benzoate, sodium benzoate, potassium benzoate, andmixtures thereof). When included in a composition of the invention, thepreservative is included at levels from about 0.0005% to about 0.5%(e.g., 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, or 0.5%) by weight basedon the total volume of the composition, though preservatives may bepresent in lower or higher concentrations.

Antioxidants

One or more antioxidant agents may also be included in the compositionsto, for example, reduce exercise-induced oxidative stress. Exemplaryantioxidants include vitamin C and vitamin E; beta-carotene, lutein, orother carotenoids; cyanidin, delphinidin, malvidin, or otheranthocyanidins; apigenin, luteolin, or other flavones; hesperitin,naringenin, or other flavonones; isorhamnetin, quercetin, kaempferol orother flavonols; and epigallocatechin-3-gallate, epicatechin,thearubigins, or other flavan-3-ols.

Additional components of the compositions described herein may includeamino acids (e.g., leucine, isoleucine, lysine, methionine,phenylalanine, threonine, tryptophan, and valine), stimulants (e.g.,caffeine), emulsifying agents, carbon dioxide (e.g., to carbonate aliquid composition), stabilizers, humectants, anticaking agents, orherbal extracts.

Combination Therapies

In certain embodiments, additional therapeutic agent(s) may beadministered with compositions of the present invention for, e.g., thetreatment or amelioration of muscle cramps and/or the recovery ofmuscles from exercise fatigue. Such therapeutic agents include, forexample, muscle relaxants (e.g., diazepam) or anti-inflammatory agents(e.g., ibuprofen). When combination therapy is employed, the additionaltherapeutic agent(s) can be administered as a separate formulation ormay be combined with any of the compositions described herein.

For example, any of the compositions described herein can be used forthe treatment of nocturnal (or night) cramps. In some embodiments, thecompositions can be used in combination with one or more sleep aids.Sleep aids that can be used in combination with the compositions andmethods described herein include: antihistamines (e.g., diphenhydramineand doxylamine); benzodiazepines (e.g., estazolam (ProSom), flurazepam(Dalmane), quazepam (Doral), temazepam (Restoril), and triazolam(Halcion)); non-benzodiazepine sedative hypnotics (e.g., eszopiclone(Lunesta), zalepon (Sonata), and zolpidem (Ambien)); and melatoninreceptor agonist hypnotics (e.g., ramelteon (Rozerem). Still other sleepaids that can be used in combination with the compositions and methodsdescribed herein include: chamomile, valerian root, kava kava, lemonbalm, passionflower, lavender, St. John's Wort, melatonin, tryptophan(e.g., L-tryptophan), 5-hydroxytryptophan (5-HTP), catnip, hops,rhodiola, oatstraw, lavender, GABA, L-theanine, linden, ginseng (e.g.,Siberian ginseng), honey, nutmeg, mugwort, butterbur, rauwolfia,taumelloolch, American hellebore, quassia, tulip tree, brewer's yeast,inositol, skullcap, phosphatidylserine, calcium, magnesium, vitamin B6,vitamin B12, and pantothenic acid (B5).

Formulations and Methods of Preparing Compositions

The compositions and solutions of the present invention may beformulated as ready-to-drink beverages, concentrates (e.g., syrups), drycompositions (e.g., powders, granules, or tablets that may bereconstituted with a liquid (e.g., with water), gels, solids,semi-solids (e.g., ice cream, pudding, or yogurt), frozen liquids (e.g.,ice pops), lozenges or hard candies, dissolving strips (e.g., an ediblestrip containing pullulan and compositions of the invention), andchewing gum.

In some embodiments, the compositions may be in the form of a drypowder, granule, or tablet that may be reconstituted in a specifiedamount of a liquid. The dried components may be mixed together andmilled (e.g., to create a homogenous powder) or mixed in aqueoussolution and dried by using methods known to one of skill in the art.Dried powders or granules may be “loose” or fashioned into tablets.

The compositions described herein can be ingested, for example, by asubject before, during, or after exercise. The compositions andsolutions described herein can be ingested (e.g., through eating ordrinking) before the onset of muscle cramping, when muscle crampingbegins, any time after the onset of muscle cramping, or after musclecramping has subsided. The compositions of the solution can also beingested after exercise to accelerate nerve-muscle recovery fromexercise fatigue. When the compositions and solutions of the presentinvention are in the form of a ready-to-drink beverage, e.g., 1, 2, 4,8, 10, 12, 14, 16, 18, 20, 24, 26, 28, 30, or 32 ounces of the beveragemay be consumed as needed (e.g., once, twice, three, four, five, sixtimes per day; once per week; or once per month).

The compositions and solutions of the present invention may be preparedusing methods known to one of skill in the art. Such methods includedissolving, dispersing, or otherwise mixing all components singularly orin suitable combinations and agitating with, for example, a mechanicalstirrer until all of the ingredients have been solubilized or adequatelydispersed. Where a shelf-stable composition or solution is desired, thefinal mixture can be pasteurized, ultra-pasteurized, sterilized, orfilled aseptically at appropriate process conditions. Where required formutual stability of two or more components (for example if a componentis unstable at low pH), multiple components can be mixed shortly beforeingestion.

The compositions and solutions described herein may be bottled orpackaged in, for example, glass bottles, plastic bottles and containers(e.g., polyethylene terephthalate or foil-lined ethylene vinyl alcohol),metal cans (e.g., coated aluminum or steel), lined cardboard containers,pouches, packs, wrappers, or any other packaging known to one of skillin the art. For example, a ready-to-drink beverage can be bottled orpackaged in a unit that contains between 10-1000 mL of the beverage. Forexample, the packaging can contain 10, 20, 50, 100, 200, 300, 400, 500,600, 700, 800, 900, or 1000 mL of the beverage. Alternatively, thepackaging can contain 200.250, 330, 350, 355.375, 440, or 500 mL of thebeverage. A ready-to-drink beverage can also be bottled or packaged in aunit that contains between 1-32 fluid ounces of beverage (e.g., the unitmay contain 1, 2, 5, 6.75, 8, 8.3.8.4, 8.45, 9.6, 10, 12, 15, 15.5, 16,18.6, 20, 23, 24, or 32 fluid ounces). Where a shelf-stable compositionor solution is desired, the packaging is appropriately sterilized beforebeing filled by the pasteurized, ultra-pasteurized, or sterilizedcomposition or solution. Where required for mutual stability of two ormore components (for example if a component is unstable at low pH), thepackaging may feature multiple containers that can be mixed shortlybefore ingestion or that can be consumed serially.

EXAMPLES General Procedures TRP-Stim Solution

The solution (“TRP-Stim”) administered to the volunteers contained: abase of a 1:1 mixture of water and light karo syrup (for increasedviscosity); 0.075% of a capsicum preparation intended for human use(Clearcap Super Soluble Caspsicum, Kalsec Inc.); 1% of a cinnamonvolatile oil intended for human consumption (Aquaresin Cinnamon, Kalsecinc); and 1.5% of a ginger oleoresin intended for human use (AquaeresinGinger, Kalsec Inc).

Electromyography (EMG) Measurements of Cramps

Methods for placing stimulating electrodes on the flexor hallucis brevis(FHB) or gastroenemius muscles followed the procedures described byMinetto et al., Muscle Nerve, 40: 535-544, 2009. The active stimulationelectrode (cathode) was a 1.25″ circular mesh-backed silver patchelectrode (Reliamed) and was placed so as to produce contraction of theFHB with minimal stimulation amplitude. The stimulation referenceelectrode was a 2″ square patch electrode (Reliamed) placed on theopposite side of the foot. Cramping of the FHB was induced as describedby Minetto et al. (ibid.) using a battery-powered electrical musclestimulator (EMS-7500, Current Solutions LLC) to deliver pulses. A seriesof 180 microsecond biphasic square pulses of voltages were applied atvarious frequencies to stimulate the muscle. First, using slow (2 Hz)stimulation, the amplitude was adjusted to ˜30% more than the thresholdamplitude for eliciting strong contraction of the muscle. The muscle wasthen stimulated by a train of 180 microsecond pulses of this amplitudedelivered for 5 seconds at various frequencies. The stimulationdelivered by the stimulator also including “ramp up” and “ramp down”periods of 1 second preceding and following the main 5-sec stimulationperiod during which the amplitude of the pulses was ramped up or down toand from the final value.

It has been previously shown that susceptibility to cramping of the FHBusing similar electrical stimulation protocols is highly reproduciblewithin each subject (Minetto et al., Muscle Nerve, 37:90-100, 2008) andis correlated with susceptibility to “ordinary muscle cramps” (Miller etal., Muscle Nerve, 39:364-368, 2009).

Cramping was quantified by making EMG recordings from the belly of theFHB. Two external EMG recording electrodes (Vermed SilveRest) wereplaced along the belly of the FHB. The differential voltage relative toa third ground electrode placed at the ankle was amplified, digitized,and saved to computer using an I-330-C2+ EMG unit with PhysioLabsoftware (J&J Engineering. Poulsbo, Wash.). The raw wide-band EMG signal(10-400 Hz) was processed by being rectified and integrated to providethe area under the curve (RMS). The duration of cramp was quantified bythe time required for the RMS EMG to return to an amplitude of 3standard deviations above the baseline value. This correlated well withduration of the cramp as observed by the return to the toe to restingposition.

Recordings of cramps in calf muscles (medial gastroenemius) were madeusing similar procedures, with placement of stimulation and recordingelectrodes following that by Minetto et al., Muscle Nerve, 40:535-544,2009. The amplitude of stimulation by a single 180 microsecond biphasicsquare pulses was adjusted to be ˜30% of the amplitude required formaximal contraction of the muscle. After a short period of slowstimulation (2 Hz), the frequency of stimulation was ramped up to 22-24Hz over ˜5 seconds and held at this frequency for an additional 5seconds before terminating the stimulation. This protocol reliablyinduced cramping of 30-90 seconds.

Assay of Activation of Rat Sensory Neurons Methods to monitor activationof primary sensory neurons isolated from the trigeminal ganglion of ratsfollowed those published by Park et al., Journal of BiologicalChemistry, 281:17304-17311, 2006). Cells isolated from rat trigeminalganglia were loaded with the fluorescent calcium indicator Fura-2AM(Fura-2-acetoxymethyl ester), and increases in intracellular calciumreflecting activation of the neurons were measured as an increase inFura-2 fluorescence as measured by digital video micro-fluorometry withan intensified CCD camera. The same capsicum extract, cinnamon extract,and ginger extract used in the TRP-Stim beverage were applied to theneurons after being diluted in balanced salt solution (in mM: 145 NaCl.5 KC, 2 CaCl₂, 1 MgCl₂, 10 HEPES, and 10 glucose) which perfused theneurons. Caspicum extract was applied at a dilution of 1/800,000,cinnamon extract at a dilution of 1/5,000, and ginger extract at adilution of 1/12,000. In some experiments the calcium ionophoreionomycin was added following the tests with extracts to produce a largeentry of calcium as an index of the maximal possible signal,illustrating the strength of activation by the heavily diluted extracts.

Example 1: Activation of Rat Sensory Neurons by Capsicum, Cinnamon, andGinger Extracts

FIG. 1 shows graphs from six sensory neurons isolated from thetrigeminal ganglia of rats, illustrating their activation by thecapsicum, cinnamon, and ginger extracts that were used in the humanexperiments. Activation was quantified as an increase in intracellularfree calcium, monitored by a fluorescent calcium indicator. Extractswere diluted into normal extracellular saline (Tyrode's solution) andwere tested at lower concentrations than used in the beverage, takingaccount that concentrations present at nerve endings in mouth,esophagus, or stomach are expected to be lower than the beverage as aresult of dilution into mucosa and interstitial fluid. All threeextracts were capable of activating individual neurons when applied atconcentrations 50-fold to 15,000-fold lower than used in the beverage.Each trace shows a record from a different neuron, illustrating thatsome neurons could be activated by each of the extracts and that thestrength of activation by each extract varied among particular neurons.These records illustrate that each agent is capable of acting alone toactivate some neurons and that a combination of agents can producestronger activation of a larger fraction of neurons. Further, the bottomtwo records show that there can be strongly synergistic activation ofneurons by the capsicum extract and the ginger extract when applied incombination.

Example 2: Effect of TRP-Stim Administration to Human Subjects

The in vitro data of Example 1 show that each individual component ofthe TRP-Stim solution by itself was capable of activating sensoryneurons. Consistent with this, human experiments showed the efficacy ofa beverage with capsicum alone (ClearCap capsicum at 1/2000 dilution) toinhibit cramping, achieved within 5 minutes.

The in vitro data also show that combinations of channel activators cannot only show the desired activity, but can also provide synergisticeffects. The following experiments, illustrated by FIGS. 2-8, show cramprelief by the administration of a uniform beverage composition designedfor maximal TRP stimulation containing capsicum, cinnamon extract, andginger extract, and where the physiological effects were monitored byEMG recording.

FIGS. 2-8 are graphs of EMG recordings of muscle contractions in sevenhuman volunteers (four females and three males) that show the efficacyin preventing and treating cramps of ingesting 50 mL of a solutiondesigned to stimulate TRPV1 and TRPA1 receptors in the mouth, esophagus,and stomach. Muscle cramps were induced by brief stimulation of toe orcalf muscles (FIGS. 2-7) or occurred spontaneously (FIG. 8). Afterrecording cramping in control, subjects drank 50 mL of the TRP-Stimsolution containing capsaicin and capsaicinoids (TRPV1 agonists),cinnamaldehyde (TRPA1 agonist), and gingerols (TRPA1 and TRPV1agonists). After ingestion of the solution, subjects were tested formuscle cramping using the same procedures as in control at times rangingfrom 4 minutes to 11 hours after ingestion.

Eight human volunteers were tested using the TRP-Stim beverage. Seven ofthe eight showed a complete abolition or dramatic reduction in crampingfollowing ingestion of the beverage (FIGS. 2-8). The effect wastypically complete within 4-15 minutes and lasted for 2½ to 4 hours indifferent subjects. An eighth subject showed cramping of the FHB thatwas not dramatically affected by the TRP-Stim beverage. The cramping inthis subject was of much lower EMG amplitude that the other subjects andappeared to involve repetitive contraction of only a few motor units.

FIG. 2 is a graph showing the effect of the TRP-Stim beverage oncramping of the flexor hallucis brevis of Subject A. Under controlconditions, cramping was reliably induced by stimulating the muscleusing an electrical muscle stimulator (EMS-7500, Current Solutions LLC)placed with external electrodes for FHB stimulation. Muscle activity wasrecorded using external electrodes placed over the belly of the muscleattached to an EMG amplifier (J&J Engineering I-330C2+). In control,stimulation using 180 microsecond biphasic pulses delivered at 18 Hz for5 seconds reliably and reproducibly produced cramping of the muscle,which was evident by EMG activity continuing after the cessation ofstimulation. After ingestion of the TRP-Stim beverage, cramping was verybrief after 11 minutes and essentially absent at tests at 20 minutes and2% hours after ingestion.

FIG. 3 is a graph showing the effect of the TRP-Stim beverage oncramping of the flexor hallucis brevis of a second subject. Undercontrol conditions, cramping was induced by stimulation at 10 Hz for 5seconds (180 microsecond pulses, amplitude set to ˜30% higher thanthreshold for muscle contraction), and a longer cramp was induced byincreasing the frequency to 12 Hz. In recordings beginning 12 minutesafter ingestion of the TRP-Stim beverage, stimulation at 10 Hz or 12 Hzproduced essentially no cramping, and increasing the frequency ofstimulation to 14 Hz also did not induce cramping. The dramaticreduction in cramping was still present 4 hours later in this subject.

FIG. 4 is a graph showing the effect of the TRP-Stim beverage oncramping of the flexor hallucis brevis of a third subject tested overlonger times. Under control conditions, a cramp lasting 58 seconds wasinduced by stimulation at 18 Hz for 5 seconds (180 microsecond pulses,amplitude set to ˜30% higher than threshold for muscle contraction).After ingestion of the TRP-Stim beverage, the duration of the cramp wasreduced to 27 seconds after 8 minutes and to 8 seconds after 15 minutes.Cramping was abolished after 20 minutes and in a test after 2 hours. Intests 11 hours after ingestion, reliable cramping had returned. Afterthe subject again drank 50 mL of the TRP-Stim beverage, cramping wascompletely abolished in tests beginning after 10 minutes.

FIG. 5 is a graph showing the effect of the TRP-Stim beverage oncramping of the flexor hallucis brevis of a fourth subject. This subjecthad engaged in strenuous exercise (triathlon) four hours earlier and wasexperiencing muscle twitchiness. This subject had an unusually lowfrequency threshold (8 Hz) for induction of cramping in the FHB muscle,and the resulting cramps were unusually long (172 seconds after 8 Hzstimulation and 222 seconds after 10 Hz stimulation). Cramping wascompletely gone in tests starting 13 minutes after ingestion of theTRP-Stim beverage, even when increasing the stimulation frequency to 12Hz. Cramping was still abolished 3 hours later. After 4 hours, crampingreturned with an increased frequency threshold (10 Hz) and shortercramps than in control. After the subject again drank 50 mL of theTRP-Stim beverage, cramping was again completely abolished.

FIG. 6 is a graph showing the effect of the TRP-Stim beverage oncramping of the gastroenemius (calf) muscle of a fifth subject. Themuscle was stimulated by a protocol ramping the frequency of stimulationfrom 2 Hz to 28 Hz (180 microsecond pulses, amplitude set to ˜30% higherthan threshold for muscle contraction). After cessation of stimulation,the muscle went into a prolonged cramp lasting 59 seconds. In a test 3minutes after ingestion of 50 mL of TRP-Stim, cramping was abolished.

FIG. 7 is a graph showing the effect of the TRP-Stim beverage oncramping of the gastroenemius (calf) muscle of a sixth subject. Themuscle was stimulated by a protocol ramping the frequency of stimulationfrom 2 Hz to 24 Hz (180 microsecond pulses, amplitude set to ˜30% higherthan threshold for muscle contraction). After cessation of stimulation,the muscle went into a prolonged cramp lasting 96 seconds. In a test 4minutes after ingestion of 50 mL of TRP-Stim, cramping was abolished.Cramping was still abolished in a test conducted 40 minutes later.

FIG. 8 is a graph showing the effect of the TRP-Stim beverage oncramping of an FHB muscle in a seventh subject, who experiencedspontaneous cramping induced by pointing her toe. In control conditions,voluntary toe flexes lasting ˜5 seconds reliably produced cramping ofthe FHB lasting 5-8 seconds in different trials. Ten minutes after thesubject ingested 50 mL of the TRP-Stim beverage, cramping was abolished.

OTHER EMBODIMENTS

From the foregoing description, it is apparent that variations andmodifications may be made to the invention described herein to adopt itto various usages and conditions. Such embodiments are also within thescope of the following claims.

All publications, patent applications, and patents mentioned in thisspecification are herein incorporated by reference to the same extent asif each independent publication, patent application, or patent wasspecifically and individually indicated to be incorporated by reference.

What is claimed is: 1-74. (canceled)
 75. A method of treating amenstrual cramp in a subject in need thereof, the method comprisingorally administering to the subject a liquid composition comprisingcapsaicinoid, cinnamaldehyde, a gingerol, and an excipient, wherein theliquid composition comprises 0.001% to 1% weight/weight (w/w)capsaicinoids per weight of the liquid composition or 0.0001-0.01 mgs oftotal capsaicinoids/mL of the liquid composition and a total of 0.001%to 1% weight/weight (w/w) of a gingerol per weight of the liquidcomposition.
 76. The method of claim 75, wherein the liquid formulationis administered prior to, during, or after the menstrual cramp.
 77. Themethod of claim 75, wherein the liquid formulation comprises a beverageor a spray.
 78. The method of claim 75, wherein the liquid formulationcomprises a beverage.
 79. The method of claim 75, wherein thecapsaicinoid comprises capsaicin.
 80. The method of claim 75, whereinthe liquid composition further comprises an electrolyte.
 81. The methodof claim 75, wherein the excipient comprises a disintegrant, a binder, asurfactant, an emulsifier, a viscosity modifier, a lubricant, asweetener, a pH-adjusting agent, a preservative, a flavoring agent, acoloring agent, or an antioxidant.
 82. The method of claim 75, whereinthe excipient is selected from gellan gum, carob bean gum, locust beangum, carrageenan, alginates, agar, guar gum, xanthan gum, carboxymethylcellulose, clear starch, pectin, gelatin, cornstarch, katakuri starch,potato starch, and gum arabic.
 83. The method of claim 75, wherein theexcipient comprises a sweetener selected from high fructose corn syrup,mannose, maltose, glucose polymers, sucrose, glucose, dextrose, lactose,galactose, fructose, polysaccharides, rice syrup, honey, saccharin,cyclamates, acetosulfam, sorbitol, sucralose, xylitol, erythritol,Stevia extract, L-aspartyl-L-phenyl-alanine ester, L-aspartyl-D-alaninealkyl amides, L-aspartyl-L-1-hydroxymethylalkaneamide, andL-aspartyl-1-hydroxyethylalkaneamide.
 84. The method of claim 75,wherein the excipient comprises a pH-adjusting agent selected fromhydrochloric acid, citric acid, sodium hydrogen carbonate, potassiumhydroxide, sodium hydroxide, and sodium carbonate.
 85. The method ofclaim 75, wherein the excipient comprises a preservative selected fromsorbic acid, benzoic acid, sodium benzoate, calcium benzoate, potassiumbenzoate, potassium sorbate, calcium sorbate, and sodium sorbate. 86.The method of claim 75, wherein the excipient comprises a flavoringagent selected from almond oil, amaretto oil, anethole, anise oil,benzaldehyde, blackberry, black walnut oil, blueberry, caraway, carawayoil, cardamom oil, cardamom seed, cherry juice, cherry syrup, cinnamon,cinnamon oil, cinnamon water, citric acid, citric acid syrup, clove oil,cocoa, coriander oil, dextrose, eriodictyon, ethyl acetate, ethylvanillin, fennel oil, ginger, glucose, glycerin, glycyrrhiza, grape,honey, lavender oil, lemon oil, lime, mannitol, methyl salicylate,myristica oil, orange oil, orange peel, orange syrup, peppermint,peppermint oil, peppermint water, phenylethyl alcohol, pineapple,raspberry juice, raspberry syrup, rosemary oil, rose oil, rose water,sarsaparilla syrup, sorbitol, spearmint, spearmint oil, strawberry,sucrose, thyme oil, tolu balsam, vanilla, vanillin, and wild cherrysyrup.